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QP99  .W87  1912      A  study  of  the  surfa 


v7oodward. 

A  Study  of  the  Surface  Tension  of  Blood  oerum 
by  the  Drop  Weight  Method. 


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A  Study  of  the  Surface  Tension 

of    Blood    Serum   by   the 

Drop  Weight  Method 


DISSERTATION 


SUBMITTED    IN    PARTIAL    FULFILMENT   OF   THE    REQUIRE- 
MENTS FOR  THE  DEGREE  OF  DOCTOR  OF  PHILOSOPHY 
IN  THE  FACULTY  OF  PURE  SCIENCE  IN  COLUMBIA 
UNIVERSITY   IN  THE  CITY   OF  NEW  YORK 


BY 

HAROLD  E.  WOODWARD,  B.  A. 

NEW  YORK  CITY 

1912 


WORCESTER,  MASS. 
Charles  W.  Bukbamk  &  Co.,  Priater« 


A  Study  of  the  Surface  Tension 

of    Blood    Serum    by   the 

Drop  Weight  Method 


DISSERTATION 

SUBMITTED    IN    PARTIAL    FULFILMENT    OF    THE    REQUIRE- 
MENTS FOR  THE   DEGREE  OF  DOCTOR  OF  PHILOSOPHY 
IN  THE  FACULTY  OF  PURE  SCIENCE  IN  COLUMBIA 
UNIVERSITY    IN   THE   CITY   OF   NEW  YORK 


BY 

HAROLD   E.  WOODWARD,  B.  A. 

NEW  YORK  CITY 

1912 


WORCESTER,    MASS. 
Charles  W.  Bureank  &  Co.,  Pr 


ACKNOWLEDGMENT 

This  investigation  was  suggested  by  Professor  J.  L. 
R.  Morgan,  and  carried  out  under  his  directions.  I  wish 
to  express  my  appreciation  of  the  assistance  and  en- 
couragement I  have  received  from  him. 

Professor  W.  J.  Gies  directed  the  experiments  on  dog 
serum,  and  I  am  very  grateful  to  him  for  the  interest  he 
has  taken  in  the  work  and  the  assistance  he  has  given  me. 

I  am  also  deeply  indetted  to  Dr.  Bailey,  Dr.  Hopkins 
and  Dr.  Smith,  of  St.  Lukes  Hospital,  for  valuable  ma- 
terial aid  and  advice,  and  to  Dr.  Butterfield  and  Dr.  Bron- 
fenbrenner,  of  the  Rockefeller  Institute,  and  Dr.  Warren, 
of  Roosevelt  Hospital. 

Harold  E.  Woodward. 

Laboratorv  of  Physical  Chemistry, 
Havemeyer  Hali,,  Columbia  University. 


CONTENTS 

Introduction  and  Object  of  the  Investigation 5 

Apparatus  and  Method 6 

Dog  Blood  Serum 11 

Human  Serum    15 

Comparison  of  Animal  Sera   17 

Meiostagmin  Reaction 20 

Summary 27 

Vita 28 


A  Study  of  the  Surface  Tension 

of    Blood    Serum    by   the 

Drop  Weight  Method 


Ascoli  and  Izar*  have  shown  that  the  immunizing 
reaction  in  pathological  blood  serum  is  accompanied  by  a 
lowering  of  the  surface  tension  of  the  serum.  They  used 
Traube's  stalagmometer,  working  at  room  temperature, 
and  observed  the  change  in  the  number  of  drops  in  a 
given  volume  of  the  serum  after  the  immunity  reaction 
had  taken  place.  It  stemd  advisable  to  confirm  this 
change,  which  they  cald  the  Meiostagmin  reaction,  by  the 
more  accurate  drop  weight  apparatus  of  Morgan  t  At  the 
same  time  it  was  thought  that  an  accurate  study  of  the 
surface  tension  of  normal  and  pathological  blood  serum 
might  be  valuable,  and  that  there  might  be  a  normal  value 
for  the  surface  tension  of  the  blood  serum  of  all  mammals. 

In  mesuring  surface  tension  by  such  methods  as  ca- 
pillary rise  or  the  drop  method  (stalagmometer),  the  den- 
sity of  the  liquid  must  be  used  to  change  the  height  of  the 
liquid  in  the  capillary  tube  or  the  drop  volume  to  surface 
tension.  But  since  drop  weights  from  any  one  tip  are 
proportional  to  surface  tensions,  for  any  liquid  and  any 
temperature,  it  is  only  necessary  to  get  the  weight  of  the 
falling  drop,  thus  avoiding  the  error  and  difSculty  of 
determining  densities.      From  drop  weights  then  we  have 

7  =  weight  X  constant 

This  work,  which  is  necessarily  rather  fragmentary, 
is  divided  into  the  following  parts, — 


*Juhnke,  Interstate  Medical  Journal,  18,  233,  (Feb.  igii) 
tMorgan,  Jour.  Amer.  Chem.  Sac.  32,  349  (191 1) 


6 

I     Surface  tension  of  blood  serum. 

A —     Dog  serum;  the  condition  of  the  dog  being 

controld. 
B —     Human  serum,  normal  and  pathological. 
C —      Comparison  of  different  animal  sera. 

II  The  Meiostagmin  reaction;  the  decrease  in  surface 
tension  accompanying  the  union  of  antibody 
and  antigen. 

APPARATUS  AND  METHOD 

The  apparatus  used  is  the  same  as  described  by  Mor- 
gan.* For  mesuring  the  drop  weight  of  serum  the  appara- 
tus was  allowd  to  remain  in  a  constant  temperature  bath 
regulated  at  37°C.  (body  temperature)  for  nearly  half  an 
hour,  then  a  drop  of  the  serum  was  puld  over  and  left 
hanging  for  five  minutes  in  order  that  the  air  in  the  weigh- 
ing vessel  should  be  saturated  with  its  vapor.  This  drop 
was  then  carefully  forced  back  into  the  supply  vessel, 
which  containd  about  5  cc.  of  serum,  and  a  fresh  drop  was 
slowly  puld  over  and  allowd  to  fall  of  its  own  weight. 
The  weighing  vessel  was  then  weighd  as  usual  after  con- 
densing the  vapor  in  it  by  water  at  room  temperature  for 
one  minute.  Next  a  blank  was  run  in  the  same  way,  ex- 
cept that  the  drop  was  not  allowd  to  fall,  and  by  sub- 
tracting this  weight  from  the  other  the  correct  weight  of 
one  drop  of  the  serum  was  obtaind. 

The  determinations  in  the  Meiostagmin  reaction  were 
made  at  o°C.  as  there  is  almost  no  evaporation  at  that 
temperature  and  in  order  that  no  reaction  should  go  on  in 
the  solution  during  the  determination.  Five  drops  were 
taken  and  weighd,  and  no  blanks  were  necessary. 

As  in  the  work  on  water  and  solutions,  the  tip  had  to 
be  cleand  after  each  determination  or  the  results  would  be 

*Loc.    cit. 


too  low.  The  cleaning  was  best  done  by  alkalin  perman- 
ganate solution  followed  by  chromic  acid  in  diluted  sul- 
furic acid,  then  the  tip  was  washt  with  distild  water  and 
dried  by  suction. 

Standardization  of  Tip 

One  tip  was  standardized  on  benzene  at  30°.     The  con- 
stant is  found  by  means  of  a  modified  Ramsay  and  Shields 


(t)" 


equation,  K  = 

t7— t— 6 

in  which  W^drop  weight  in  milligrams,  M^yS,  d=. 86824, 
andt<r=288  5.  Surface  tensions  may  be  found  from  the 
ratio 

K' 
W:  7   ::   K   :   K'     or     7=   —   W 

y  will  be  in  dynes  per  centimeter,  and  K'  is  the  surface 
tension  constant  from  the  Ramsay  and  Shields  formula, 
K'=  2.1148.* 

Tip  I  gave  the  following  results  with  benzene  at  30°. 

Vessel  -j-       Vessel  + 


30  drops 

5  drops 

11.7112 

10.9695 

11.7113 

10  9694 

25  drops 

=  0.74182  gram. 

11.7114 

10  9697 

11.7113 

10.9695 

I  drop 

=  0.0296728 

11.7114 

10.9693 

11.7112 

10.96948 

=  29.67  mg. 

11.71130 

K  =  2.3572 

2.1148 

2  3572 

W  =  0.8972 

W 

*Morgan  and  McAfee,  Jour.    Timer.  Chem.  Soc.  33,  1275.  (1911) 


diameter  of  tip*   ^  — =-^^ =  5.5S0  mm. 

,4224 

Two  other  tips  were  standardized  on  distild  water  at 
37°.     The  surface  tension  of  water,  from 

7  =  75.872  —  0.1547/  —  0.000222/^   is  69.844  at  37° 
Tip  II  gave  results  as  follows, 

Vessel  -{-       Vessel  -{- 
30   drops       5  drops 

13.2202  11.3082  25  drops  ^=  1. 91208     grams. 

13.2207  11.3085 

13.2199  11.3078  I     drop    =  0.0764832 

13.2202  II  3082 

13  22025  II. 30817     _  =  76.483  mg. 
69  844 


76.483 


W  =  0.91319  W 


2.IT48  „    ■ 

K  = ^—      =  2.3158 

■91319 

,    .  2  3158 

diameter  of  tip  =  — = — = —  =:  5.483  mm. 
.4224 

Results  on  Tip  III  were 

Vessel  -\-       Vessel  + 
30    drops       5  drops 

12.9084  II. 0109  25  drops  =  1.8976 

12  9085  II  0108 

12  9082      II  0106      I  drop  =  0.075904 

12.90837     1 1. 01077  =  75  904  mg. 
69  844 


75  904 


W  =  0.92016  W 


"Morgan  and  Cann,  Jour,  iinier.  Chem.  Soc.  33,   1060  (1911) 


.92016 


=  2.2983 


,,  ,     .  2.2q83 

diameter  of  tip  = =  5.441  mm. 

.4224 

In  order  to  test  the  tips  with  thick  viscous  liquids 
somewhat  like  serum,  the  drop  weights  of  glycol  and  sugar 
solutions  were  first  tried. 

Glycol  has  practically  no  vapor  tension  at  ordinary 
temperatures,  (b.  p.  200°)  so  10  drops  were  weighd  without 
any  blank.  At  0°  glycol  is  so  viscous  that  it  took  over 
thirty  minutes  to  get  the  ten  drops,  while  the  same  num- 
ber of  drops  of  water  are  generally  dropt  in  less  than  five 
minutes. 

Results  on  glycol   (CH^OH)^ 


Temp.  Tip 

10  drops 

Average 

I  drop 

Crit. 

grams 

mgs. 

Temp. 

0          I 

0.5265 

0.5266 

0.5273 

0.5269   0.52683   52-683   329.5   47.26 
0.5270 
0.5268 
30  I    0.5051 
0.5053 

0.5047   0.50504   50.504   350.0   45.31 
0.5051 
0.5050 

55       III        0.4728 
0.4733 

0.4734         0.4731  47-310         365.9         43.53 

04729 

The  values  for  surface  tension  give,  by  least  squares, 
the  formula,  7  =  47.277  —  0.0675/.  Walden's*  formula, 
from  capillary  rise  is 

7  =  48.48  (i  —  0.00205/),  or  7  =  48.48  —  0.0994/. 
*Walden,  Zeits.  Phys.  Chem.  65,  143. 


10 

The  large  difference  at  low  temperatures  is  probablj^  due 
to  the  fact  that  the  liquid  is  so  viscous  at  o°  that  the  rise 
in  a  capillary  tube  can  not  be  accurately  mesured. 

Comparison  of  results 

7  =  47.277   —  0.0675^  7  =  48  48  —  0.0994/ 
o                             47-28  48.48 

30  45-25  45  50 

55  43-57  43-OI 


Tip 
III 


Sugar  solutions  at  37° 


II  0,3881         0.0780         87.63         70  86 


5  drops 

I  drop 

W 

.5  molar 

0.3819 

0.0770 

76.50 

0.3S26 

0.0768 

76  30 

0.3829 

—  .0006 

0.3825 

76.40 

I  molar 

0,3881 

0.0780 

87.63 

03882 

0.0780 

—  .0005 

77  50 

0.38815 

77  56 

1.5  molar 

0.3887 

0.0785 

77.81 

0.3894 

0.0784 
=  .0005 

77  95 

0.38905 

77.88 

2  molar 

0.3983 

0.0804 

79.81 

0.3098 

0.0803 

79-85 

70.30 


III   0.3887   0.0785   77.81   71.67 


II     0.3983    0.0804    79-81    72.90 


0.39905  -.0005    79.83 


The  method  of  least  squares  applied  to  these  results, 
with  69.84  as  the  surface  tension  of  water,  gives 
7  =  69.878  4-  0.456C  +  0.5202c'' 


11 


for  the  surface  tension  of  different  concentrations  of  sugar 
at  37^.  The  calculated  results  from  this  agree  very  well 
with  the  observd  results. 


obs. 

calc. 

diff. 

Water 

69.84 

69.88 

+0.04 

0.5  molar  sugar 

70.30 

70.24 

—0.06 

I          "           " 

70.86 

70.85 

— O.OI 

1.5       " 

71.67 

71-73 

+0.06 

2          "           " 

72.90 

72.87 

-0.03 

DOG  BLOOD  SERUM 


Except  for  the  first  three  which  were  used  for  practis, 
the  dogs  were  kept  in  special  cages  and  fed  the  regular 
diet  (15  gr.  meat,  4  gr.  cracker  meal,  3  gr.  lard  and  35  cc. 
water  per  kilogram  of  body  weight)  until  they  were  in 
normal  condition.  They  were  then  subjected  to  various 
conditions  which  a  person  might  undergo,  such  as  missing 
meals,  having  extra  food,  loss  of  blood,  action  of  cathartic, 
etc.,  and  they  were  then  bled  in  order  to  see  if  the  surface 
tension  of  the  blood  serum  would  change  under  such  con- 
ditions. 

At  first  a  few  mesurements  made  on  defibrinated  blood 
and  contrifuged  serum,  to  see  if  the  surface  tension  was 
the  same  in  either  case.  There  seemed  to  be  almost  no 
difference,  altho  a  French  investigator  reported  a  differ- 
ence of  over  2%.     Results  on  one  dog  were,  with  Tip  I. 


Blood 
5  drops 

0.2487 
0.2479 


W 
49.66 


44.6 


Serum 
2  drops 

0.0991 
0.0992 
0.0993 
0.0989 


W 


49-56 


44-5 


12 


As  the  serum  was  easier  to  use,  and  the  blood  decomposed 
rather  easily,  the  serum  was  used  alone. 

A  comparison  was  then  tried  between  centrifuged  and 
clotted  serum,  to  see  if  the  method  of  getting  the  serum 
made  any  difierence  Serum  from  one  dog  gave  on  Tip  III. 


Centrifuged 
I  drop  W 


0.0502 
0.0502 


49-7 


45-7 


Clotted 
I  drop 

0.0499 
00497 


W 


49-3 


45-4 


— .0005  —.0005 

There  is  thus  a  little  more  than  half  of  one  percent 
difference,  but  as  the  clotted  serum  remaind  clear  longer 
than  the  centrifuged,  the  blood  was  allowd  to  clot  after 
No.  7. 


No. 


Tip 
I 


Experiments  on   Dogs 

Dog  I  drop        W  7 

I  Fern.    0.0991 

0.0992     49.56     44.5 

0.0993 

0.0989 

[      "       0.2560 

0.2559     51-19     45-9 

0.2560 

0.2558 


Remarks 


2  Male 


4   n 


5     III     4  Fem. 


0.2757 
0.2765 
0.2760 

0.2759 

0.0502 
0.0499 

0.0503 
0.0500 
0.0503 


55-2       49-5 


49.6       45.7 


49.6       45.7 


Dog  did  not  eat  or 
urinate  for  several 
days. 


13 

6  III     5  Male    o  0494     48.9       45.0 

00493 

7  III     5      "        0.0502     49  7       45  7     Centrifuged 

00502 

0.0499     49  3       45  4     Clotted 
0.0497 

8  III     6      "        0.0500     49.4       45.5 

0.0498 

9  III     6      "        00498     49.3       45.4     Same  dog,  next  day 

0.0498 

10  II      4  Fern.    0,0496     49  I       44.8     Extra  meal 
III  0.0490     48.0       44.7 

0.0491 

11  II       4      "        0.0496     49  I       44  8     Extra  meal 
III  o  0492     48.7       44.8 

0.0493 
0.0491 

12  III     5  Male    00499     494       45.5     No  food  day  before 

o  0499  bleeding 

o  0505     49  9       45.9     No    food     for    two 
00503  days 

0.0499     49.3       54  4     Extra  water 
0.0497 

0.0508     50.3       45.9     Did  not  eat  for  se- 
0.0504     49.9       45.9     veral  days 

16  III  8   "   0.0500 

0.0502  49.7   45.7  10  gr.  salt 
0.0502 

17  III  9  Fem.  0.04931 

0.04930  48.8       44.9     10  gr.  sugar 
0.04924 


13  III 

6 

14   III 

6 

15    n 

7 

III 

14 


0.04980 

0.04984 

49-3 

45-4 

Doseof  Magnesium 

0.04968 

sulfate 

0.0502 

49-6 

45-6 

0.0501 

0.0501 

0,0504 

49.8 

45.8 

Same  dog  next  day 

0.0503 

0.0503 

18     III     9 


19     III  10 


III 


NOTE: — In  No.  i  two  drops  were  weighd,  and  in  No.  2  and  3  five 
drops  were  weighd  in  a  vessel  containing  some  serum.  In  the  others 
one  drop  was  weighd  in  an  empty  vessel  and  the  weight  of  the  vapor, 
as  a  result  of  several  determinations  was  taken  as  0.5  mg. 

The  first  eight  experiments  show  that  the  normal 
surface  tension  of  dog  blood  is  about  45.5  dynes  per  centi- 
meter. The  first  three  vary  from  this  more  than  the  others 
because  the  dogs  had  not  been  kept  in  condition. 

Experiment  9  was  to  see  if  the  loss  of  blood  had  any 
effect  on  the  surface  tension.  The  dog  was  bled  about  as 
far  as  was  safe  each  day  (No.  8  and  9),  and  the  surface 
tension  was  about  the  same  each  day.  This  experiment 
was  repeated  in  No.  19  and  20,  except  that  the  two  bleed- 
ings were  about  19  hours  apart  insted  of  26  hours.  In  this 
shorter  time  the  surface  tension  had  not  quite  reached  its 
normal  value,  for  the  blood  was  still  rather  diluted. 

In  No.  10  and  11  the  dog  was  given  an  extra  meal 
several  hours  before  the  blood  was  taken.  The  decrease 
of  about  1.5%  shows  the  effect  of  the  products  of  digestion, 
especially  fat,  on  the  surface  tension  of  the  blood. 

In  experiments  12  and  13  the  dog  starvd  one  and  two 
days,  respectively,  before  the  blood  was  taken.  The  re- 
sult in  No.  13  shows  a  rise  of  about  1%  in  surface  tension 
as  the  blood  becomes  poorer  in  the  products  of  digestion. 
The  dog  in  No.  15  refused  to  eat  for  several  days,  and  the 
surface  tension  in  this  case  confirms  the  other  result. 


15 

500  cc.  of  water  given  two  hours  before  the  bleeding 
in  No.  14  seetnd  to  have  no  effect  on  the  surface  tension, 
and  a  good  dose  of  magnesium  sulfate,  in  No.  18,  which 
would  remove  water  from  the  system,  did  not  seem  to 
cause  any  change. 

In  experiment  16  the  last  meal  before  bleeding  con- 
tained a  large  amount  of  salt,  and  the  surface  tension  was 
raisd  a  little,  as  it  should  be  if  much  salt  was  taken  up 
by  the  blood.  The  effect  of  sugar  in  the  diet  is  shown  in 
No.  17,  where  there  is  a  decided  decrease.  This  may  be 
because  glucose   lowers    the    surface  tension  of  solutions. 

HUMAN   SERUM 

The  results  on  human  serum  are  not  as  satisfactory  as 
those  on  dog  serum,  as  it  has  been  shown  that  the  amount 
and  kind  of  food  eaten  has  an  effect  on  the  surtace  tension 
of  the  serum,  and  the  conditions  governing  the  human 
subjects  were  not  regulated,  as  was  the  case  with  the  dogs. 
Most  of  this  serum  was  obtaind  from  St.  lyuke's  and  Sloan 
Hospitals,  and  some  was  given  by  fellow  workers  in  the 
laboratory. 

Results  on  human  serum. 


No. 

Tip 

I  drop  gr. 

W  mg. 

7 

Remj 

I 

I 

0.0499 

0.0500 

49-5 

44'5 

0.0500 

2 

I 

0.2577 
0.2578 

51-5 

46.2 

Placental 

3 

II 

0  0507 

50-3 

45'9 

" 

4 

II 

O.0511 

0.0514 

50.8 

46.4 

" 

0.0512 

16 


5 

11 

0.0502 

49.8 

45-5 

" 

0.0503 

6 

II 

0.0520 

0.0521 

51.6 

47.1 

Mict 

0.0520 

7 

II 

0.0530 
0.0532  ■ 

52.6 

48.0 

Kidney 

8 

III 

0.0563 
0.0565 

55-9 

51-4 

Chronic  nephritis 

9 

III 

0.0517 
0.0519 

51-3 

47.1 

Same,  later 

lO 

I 

0.0499 
0.0501 

49-5 

45-1 

"          " 

II 

III 

0-0535 
0.0535 

53-0 

48.8 

IvOC.  atax. 

12 

III 

0  0492 

48.7 

44.8 

Apoplexy 

II 

00495 

49.0 

44-7 

13 

III 

0.0489 

0.0491 

48.6 

44-7 

J.  S.  B. 

0.0492 

14 

III 

0.04876 

0.04856 

48.1 

44-3 

" 

0.04863 

15 

III 

0.0499 
0  0498 

49-35 

45-4 

F.  R.  K. 

16 

III 

0.0506 

0.0506 

50.1 

46.1 

H.  E.  W. 

0  05068 

17 

III 

0  04968 

0.04952 

49.1 

45-2 

" 

0  04964 

17 

NOTE: — In  all  of  these,  except  No.  2  in  which  five  drops  were 
weighd,  a  single  drop  was  weighd  in  the  empty  vessel  and  the  weight 
of  the  vapor,  as  a  result  of  several  determinations,  was  taken  as  0.5  mg. 

Of  these,  No.  1—5  and  13 — 17  are  probably  quite  nor- 
mal, and  give  an  average  value  of  45.4  dynes  as  the  sur- 
face tension  of  human  serum,  but  most  of  them  vary  con- 
siderably from  this  because  the  conditions  were  not 
controld  as  was  the  case  with  the  dogs.  The  placental 
sera  (No.  2 — 5)  are  rather  high  (average  =  46.0),  pro- 
bably because  the  patients  had  not  been  eating  as  much 
as  the  others  (average  =  450).  The  blood  in  No.  13  and 
14  was  obtained  in  the  afternoon  while  it  was  taking  up 
the  digested  food,  which  is  the  reason  for  the  low  value. 
No.  15  was  obtained  about  an  hour  after  eating,  before 
the  blood  could  absorb  any  food.  No.  16  was  taken  at 
noon — about  eighteen  hours  after  eating,  since  nothing 
was  eaten  that  morning — while  a  few  days  later  No.  17 
was  taken  about  four  hours  after  eating,  thus  showing  the 
great  variation  in  one  individual  at  different  times  of  day. 

Several  of  the  pathological  sera  show  a  higher  surface 
tension  than  would  be  expected  even  from  a  low  diet.  In 
the  case  of  the  kidney  trouble  and  nephritis  this  is  pro- 
bably due  to  retention  of  salt,  and  in  the  latter  it  accom- 
panies a  high  blood  pressure. 


COMPARISON  OF   DIFFERENT 
ANIMAL  SERA 

The  blood  serum  of  horse,  rabbit,  guinea-pig  and 
sheep  was  obtaind  from  Rockefeller  Institute  and  St.  lyuke's 
Hospital,  for  comparison  with  dog  and  human  serum.  The 
results  do  not  agree  as  closely  as  they  did  on  dog  serum, 
since  the  condition  of  these  animals  was  not  governd 
quite  as  carefully  as  with  the  dogs. 


18 

Results  on  animal  serum 

No.       Tip       Animal      i  drop  W  7 

1  III         Horse         0.0516 

00514         51.0         46.9 
0.0514 

2  "  0.0488 

0.0488         48.3         44.5 
0.04S8 

3  Rabbit         0.0496         49.1         45.2 

0.0495 

4  "  0.0506         50.1  46.1 

0.0505 

5  "  0.0521  51.6         47.5 

0.0521 
0.0521 

6  Guinea         0.0503         49.8         45.3 

pig  0.0503 

7  Sheep         0.05223       5T.7         47.6 

0.05229 

8  "  0.04922       48.8         44.9 

o  04941 

Comparison  of  surface  tensions 

Human  from     44.3  to     46.4  av.     45.4 

Dog  44.7  45.9  45.3 

Horse  44.5  46.9  45.7 

Rabbit  45.2  47.5  46.3 

Guinea  pig  45.3  45.3 

Sheep  44.9  47.6  46.2 

The  surface  tension  of  blood  serum  is  much  below 
that  of  water,  but  this  is  not  surprising  when  the  number 
of  substances  in  solution  is  considerd.     The  list  includes 


19 

neucleoprotein,  serum  globulin,  serum  albumin,  dextrose, 
fat,  enzymes,  lecethin,  cholesterol  and  esters,  gases,  color- 
ing matter  and  inorganic  salts  such  as  chlorides,  phos- 
phates, carbonates  and  sulfates  of  sodium  potassium  and 
magnesium.  The  only  ones  of  this  list  which  raise  the 
surface  tension  of  water  are  the  inorganic  salts,  while  the 
proteins,  being  colloids,  would  lower  it  somewhat  and 
emulsions  of  fat,  lecethin  and  chloresterol  lower  it  still 
more. 

The  surface  tension  of  serum  has  been  mesured  by  a 
French  investigator*  using  a  stalagmometer.  In  this 
method  the  number  of  drops  of  serum  in  a  certain  volume 
is  counted  and  compared  with  the  number  of  drops  of 
water  in  the  same  volume  of  water  as  follows — 

No.  of  drops  of  water  X  density  of  serum 

X   75  = 

No.  of  drops  of  serum 

surface  tension 

He  found  it  necessary  to  determine  the  water  "constant" 
several  times  a  day,  so  the  method  can  not  be  very  ac- 
curate. The  use  of  75  as  the  surface  tension  of  water 
shows  that  he  probably  workt  at  low  temperatures  (about 
5°C),  instead  of  body  temperature.  His  values  vary  from 
69  to  74  (almost  equal  to  pure  water),  while  a  few  deter- 
minations by  drop  weights  at  0°  gave  about  65.  He  gave 
results  for  different  dilutions  of  serum  with  water,  which 
do  not  agree  very  well.  In  some  cases  the  very  dilute  so- 
lutions gave  results  above  water,  and  in  others  they  were 
nearer  the  value  for  undiluted  serum.  The  drop  weights 
of  a  few  solutions  of  serum  were  taken,  but  no  irregular 
curves  like  those  were  found.  The  drop  weight  work  on 
solutions  was  not  continued,  as  these  did  not  seem  to  be 
any  constant  cure  for  dilution,  and  the  very  dilute  solu- 
tions decompose  too  easily.     A  knowledge  of  the  surface 


*Iscovesco,  Comp.  Rend.  Soc.  Biol-  69,  353,  70,  1166,  93,  etc. 


20 

tension  of  serum  may  be  of  importance  in  the  case  of  injec- 
tion of  salt  solution  after  loss  of  blood,  as  this  is  generally 
followd  by  a  fever,  since  the  surface  tension  of  the  injected 
salt  solution  is  higher  than  that  of  the  serum.  It  may  be 
that  a  study  of  serum  diluted  with  salt  solution  may  be  of 
importance  in  this  connection. 

Some  Italian  investigators*  have  used  the  capillary 
rise  method  on  blood  serum,  but  they  worked  at  nearly 
all  temperatues  except  body  temperature,  so  the  results 
can  not  be  compared.  Both  the  French  and  Italian 
workers  have  said  that  the  surface  tension  of  the  serum  is 
less  than  that  of  distild  water  or  isotonic  salt  solution,  and 
varies  according  to  the  species  and  the  individual.  The 
results  of  this  investigation  show  that  the  variation  in 
any  one  individual  may  be  as  great  as  between  different 
individuals,  and  the  value  for  any  individual  is  about  45 
or  46  dynes. 


MEIOSTAGMIN   REACTION 

Ascoli  and  Izar,  of  the  University  of  Pavia,  who  ori- 
ginated the  Meiostagmin  reaction,  showd  that  when  a 
specific  antigen  is  added  to  a  specific  serum,  the  surface 
tension  of  the  liquid  is  lowerd  after  the  antibody,  present  in 
the  pathological  serum,  has  bound  the  antigen,  by  incuba- 
tion for  two  hours  at  37°.  The  reaction  has  been  tried  on 
typhoid,  syfilis,  tuberculosis  and  cancer  serum,  being 
positive  in  about  90%  of  the  cases  which  are  positive  by 
standard  methods,  and  nearly  always  negative  in  negative 
cases. 

This  reaction  was  tried  on  the  drop  weight  apparatus 
in  a  series  of  syfilis  cases — about  25  being  positive  by  the 
Wassermann  and  5  negative.     The  serum  came  from  St. 

*Fano  and  Mayer,  slrcli.    di  Fisiol.  4,  165 


21 

Luke's  and  Roosevelt  Hospitals,  and  the  Wassermann  tests 
were  made  in  their  laboratories.  It  seemd  best  to  make 
the  determinations  at  o°  instead  of  at  room  temperature, 
so  that  no  reaction  would  be  going  on  while  the  determin- 
ations were  being  made,  and  water  was  used  for  diluting 
since  there  were  no  corpuscles  present, — otherwise  the  re- 
action was  carried  out  according  to  directions.  The 
reaction  does  not  seem  to  be  as  quick  as  many  such 
reactions.  When  red  blood  corpuscles,  for  example,  are 
treated  at  o°  with  an  antibody  (from  the  serum  of  an  animal 
which  has  been  immunized  against  these  corpusclse)^ 
and  the  corpuscles  are  immediately  centrifuged  away  from 
the  solution,  it  is  found  that  they  have  combind  with  all 
the  antibody  in  the  few  seconds  that  they  were  together. 
In  the  Meiostagmin  reaction  it  takes  about  two  hours  for 
complete  union  of  the  antibody  and  antigen  at  37°  and 
there  seems  to  be  almost  no  union  at  0°. 

Izar  used  alcoholic  ethereal  splenic  extract  from  a 
syfilitic  fetus  for  an  antigen,  but  in  this  work  the  antigens 
were  mostly  from  beef  heart.     The  following  were  used, — 

Antigen  I  The  acetone  insoluble  part  of  an  alcoholic 
and  ethereal  extract  of  beef  heart,  in  al- 
cohol solution.  Titrated  strength  is  .04 
cc.  to  .1  cc.  of  serum. 

"        II         An  alcoholic  extract  of  guinea  pig  heart. 
Strength  is  .06  cc.   .1  cc.  serum. 

"        III       Same  as  I.      Strength    is  .01  cc.  to  .i  cc. 
serum. 

The  solutions  were  made  up  so  that  the  serum  was  i  to  25. 

Antigen  I  0.2  cc.  II  0.3  cc.  Ill  0.05  cc. 
Water                     11. 8                        11. 7  ii-95 

Serum  0.5  0.5  0.5 

Total  12.5  12.5  12.5 


22 

Half  of  the  solution  was  put  in  the  incubator,  while 
the  other  half  was  put  in  the  appartus  and  its  surface  ten- 
sion determind  at  once.  The  weight  of  five  drops  was 
found  and  the  determination  was  repeated  to  get  a  check 
within  I  mg. — this  allowd  an  error  of  about  o.i%  and 
most  of  the  changes  were  over  0.5%. 

It  seemd  to  make  no  difference  whether  the  serum  was 
inactivated  (by  heating  to  55°  for  30  minutes)  or  not,  but 
most  of  the  samples  were  inactivated,  as  they  had  been 
used  for  the  Wassermann  test  in  which  the  serum  must 
contain  no  complement.  In  one  case  the  Meiostagmin  re- 
action was  tried  both  before  and  after  inactivation  of  the 
serum.  Inactivation  destroys  the  complement  (which  is 
present  in  all  serum),  but  leaves  the  immune  body,  and  it 
is  the  latter  which  is  necessary  for  this  reaction. 


Tip  III 

Fresh  Se: 

rum                     Antigen  III 

Before  incubation 

After  incubation 

5  drops        W               7 

5  drops           W              7 

0  3976 

0-3955 

03981         79-57         73- 

22 

0-3965         79.20         72  88 

0.39785  0.3960 

Same  Serum  after  30  minutes  heating  at  55° 

0.4072  0.4057 

0.4070    81.42    74  35  0.4047    81.04    74-OI 

o  4071  0.4052 

Change  in  both  cases  is  —0.5% 

The  Meiostagmin  reaction  works  therefore,  whether 
the  serum  is  fresh  and  contains  complement  or  whether  it 
has  lost  its  complement  either  by  keeping  it  three  or  four 
days  or  by  heating  it. 

Five  non-syfilitic  cases  were  tried,  four  being  negative 
by  the  Wassermann  test,  and  the  fifth  being  blood  serum 
of  the  author  which  was  used  as  a  control. 


23 


Tip  III  Results  on  non-syfilitic  cases 

Before  incubation  After  incubation         Change 


■51 


No.Ant. 

5  drops 

w 

I    I 

0-3753 
0.3749 
0.3751 

75.02 

2    II 

0  3577 
0.3586 
0.3581 

71.62 

3   HI 

0  4056 
0.4051 
0-40535 

81.07 

4  III 

0.4006 
0.4010 
0.4008 

80.16 

5  in 

0.3997 
0.4007 

80.04 

6536 


74.60 


73.76 


7365 


5  drops 

W 

7 

0.3757 

74.99 

68.49 

0.3742 

0.37495 

0.3593 

71.93 

65.66 

0.3600 

0-35965 

0.4048 

80.90 

74.45 

0.4043 

0.40455 

0.4008 

80.16 

73-76 

+0.3 


0.4002 


0.3999 

o  4005 
0.4002 


80.04  73.65 


The  negatives  do  not  show  more  than  0.2%  decrease, 
and  that  is  about  the  limit  of  accuracy  with  these  solu- 
tions, as  it  means  a  check  of  0.8  mg.  on  the  weight  of  five 
drops  before  and  after  incubations.  The  one  in  which  An- 
tigen II  was  used,  probably  showd  an  increase  on  account 
of  the  large  amount  of  the  alcohol  solution  which  had  to 
be  used. 

Results  on  positive  syfilis 

Before  incubation  After  incubation 


No.Ant.  5  drops 

W 

7 

5  drops 

W 

7        Change 

gr. 

mg. 

gr. 

mg. 

% 

6  I        0.7507 

75-03 

69.04 

0  7412 

74.14 

67.30      -2.5 

0.7499 

0.7416 

0.7503 


0.7414 


24 

7  0-3337  66.80  6i.oo  0.3261  65.23  59.54  —2.4 
03343  0.3262 

0.3340 

8  03635  72.67  66.87  0.3565  71.36  65.67  -18 
0-3632  0-3571 

0.36335  0.3568 

9  0.7927  79  30  72.42  0.3969  79.28  72.40    o 
0.7932  o  3959 

0.79295  o  3964 

10  II     0.3537  70.84     65.15     0.3343     66.92     6r.6o     —5.5 

0-3547  0-3350 

0-3542  0.3346 

11  03591  71.82     66.07     0.3523     70.46     6487     —1.8 
0.3591 

12  0.3575  71-42     65.72     0.3550     70.89     65.23     —08 
0-3567  0-3539 

0.3571  0.3544 

13  III  0.3783  75.80  69.75  0.3710  74.26  68.33  -2. 

0.3797  0-3716 

0.3790  0.3713 

14  0.3963  79.37  73.04  0.3885  77.76  71.55  -2. 
0-3974  0.3881 

0.3968  0.3883 

15  0.3881  77.53  71.34  0.3818  76.28  70.19  —1.6 
o  3872  0.3810 

0.3876  03814 

16  0.3955  79-o6  72.75  0.3914  78.29  72.04  —I. 
0-3951  03915 

0.3953 

17  0.3976  79.57  73.22  0.3955  79-20  72  88  -0.5 
0-3981  0.3965 

0.3978  o  3960 


25 

58     0.4070  8i.34  74-85  0.4053  81  01  74.55  —0.4 

o  4064  0.4048 

0.4057  0.4050 

19  0.4033  80.64  74-20  0.4025  80.42  74.00  —0.3 
0.4031  0.4017 

0.4032  0.4021 

20  0.3965  79.26  72.94  0.3951  79.06  72.75  —0.26 
o_3q6i_  0-3955 

0.3963  0.3953 

21  0.4046  80.96  74.50  0.4039  80.84  74-39  —0.15 
0.4050  0.4045 

0.4048  0.4042 

22  0.4027  80.60  74.17  0.4027  80.54  74-11  —0.1 
0403.-^  0.4027 

o  4030 

23  0.3909  78.28  72.03  0.3721  74.52  68-57  -4-8 
0-3919  0.3731 

0.3914  0.3726 

24  0.3949  79-03  72-72  0.3873  77.56  71.37  -1.9 
0.3954  0.3883 

0.3951  0.3878 

25  0.3985  79.64  73.28  0.3918  78.28  7203  -1.7 
o  .•^97g  0.3910 

0.3982  0.3914 

25     0.3992  79.84  73.47  0.3944  78.77  72.49  -1-3 

0-3992  0.3933 
o  3938 

27     0.4024  80.54  74-IO  0.3999  80.04  73-65  —0.6 

0.4030  0.4005 

0.4027  0.4002 


26 

28  0.3997     79-97     73-59     0.3974     79-58     73-23     -o-3 

0-3990  0.3984 

0.3993  0.3979 

NOTE: — In  No.  6  and  the  first  half  of  9,  tea  drops  were  weighd,  in- 
sted  of  five.  No.  7  and  9  were  on  Tip  II,  all  the  others  on  Tip  III.  In 
No.  23 — 28,  the  solutions  were  made  up  so  that  the  serum  was  i  volume 
in  10,  insted  of  i  in  25,  with  apparently  no  difference  in  results.  Re- 
sults on  four  other  positive  cases  which  gave  a  decided  decrease  in 
surface  tension  are  ommitted  as  the  results  were  not  checkd  very 
closely  on  account  of  the  tip  not  being  cleand  well  enough. 

In  all  but  three  of  these  there  is  a  decided  decrease  in 
surface  tension  after  incubation,  but  in  No.  9,  21  and  22 
the  change  is  no  greater  than  might  be  found  in  a  nega- 
tive case,  so  these  can  not  be  calld  positive  by  the  Meios- 
tagmin  reaction.  The  history  of  these  three  cases,  how- 
ever offers  an  explanation.  No.  9  was  a  case  in  which  the 
primary  occurd  twenty-five  years  ago,  and  it  was  cured  at 
that  time  by  the  regular  treatment.  The  Wassermann  test 
was  a  very  weak  positive,  as  titration  showd  only  two 
units  of  antibody  while  an  active  case  contains  about  eight 
or  ten  times  as  much.  No.  22  was  a  case  of  tertiary 
syfilis  in  which  the  primary  was  ten  years  ago.  It  gave  a 
positive  Wassermann  with  one  antigen  and  negative  with 
another.  '  The  Wassermann  test  was  not  tried  with  the  an- 
tigen which  was  used  in  the  Meiostagmin.  No.  21  was  a 
case  of  early  tabes  probably  on  syfilitic  basis.  The  Meio- 
stagmin reaction  is  almost  positive,  but  is  very  doubtful. 
No.  20,  which  gives  a  weak  positive  Meiostagmin,  gave  a 
doubtful  positive  Wassermann  test.  The  case  had  been 
treated  for  about  eight  months  with  mercury  and  potassium 
iodide. 

The  Meiostagmin  reaction  is  evidently  not  quite  as 
delicate  as  the  Wassermann  at  present,  tho  it  probably 
would  be  more  delicate  if  the  average  of  three  or  four  de- 
terminations were  taken  insted  of  two,    or  if  more  drops 


27 

were  weighd.  It  is  a  simpler  test  to  make,  for  it  only  re- 
quires the  serum  and  antigen,  while  the  Wassermann  re- 
quires inactivated  serum,  antigen,  guinea  pig  serum  for 
complement,  sheep  blood  corpuscles  and  rabbit  serum 
which  has  been  injected  with  sheep  corpuscles,  all  of 
which  must  be  tested  to  see  if  they  can  be  used  together. 

SUMMARY 

1.  The  surface  tension  of  the  blood  serum  in  an  in- 
dividual may  change  during  the  day,  depending  on  the 
food  that  is  being  absorbd  by  the  blood. 

2.  The  surface  tension  of  the  blood  serum  of  different 
individuals  and  different  species  is  approximately  the 
same,  if  account  is  taken  of  the  daily  change  in  any  one 
individual. 

3.  The  surface  tension  of  the  blood  serum  seems  to 
be  abnormally  high  in  certain  diseases,  especially  those  in 
which  the  kidneys  are  affected. 

4.  The  Meiostagmin  reaction  was  found  to  be  speci- 
fic in  about  87%  of  a  series  of  clinically  positive  syfilis 
cases. 

5.  The  clinically  positive  cases  of  syfilis  in  which 
the  Meiostagmin  reaction  was  not  positive  were  those  in 
which  the  Wassermann  test  was  weak  or  rather  doubtful. 


VITA 

Harold  Edward  Woodward  was  born  in  Worcester, 
Massachusetts,  July  5,  1888.  He  receivd  the  degree  of 
Bachelor  of  Arts  from  Amherst  College,  in  June,  1910. 
Since  then  he  has  been  studying  Chemistry  at  Columbia 
University,  under  the  Faculty  of  Pure  Science,  for  the 
degree  of  Doctor  of  Philosophy.  During  the  year  1910- 
191 1  he  taught  Chemistry  in  Horace  Mann  High  School, 
and  he  was  Samuel  Anthony  Goldschmidt  Fellow  in 
Chemistry  for  1911-1912. 


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